• Title/Summary/Keyword: isolation, identification

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Glycosyl glycerides from hydroponic Panax ginseng inhibited NO production in lipopolysaccharide-stimulated RAW264.7 cells

  • Cha, Byeong-Ju;Park, Ji-Hae;Shrestha, Sabina;Baek, Nam-In;Lee, Sang Min;Lee, Tae Hoon;Kim, Jiyoung;Kim, Geum-Soog;Kim, Seung-Yu;Lee, Dae-Young
    • Journal of Ginseng Research
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    • v.39 no.2
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    • pp.162-168
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    • 2015
  • Background: Although the aerial parts of hydroponic Panax ginseng are reported to contain higher contents of total ginsenosides than those of roots, the isolation and identification of active metabolites from the aerial parts of hydroponic P. ginseng have not been carried out so far. Methods: The aerial parts of hydroponic P. ginseng were applied on repeated silica gel and octadecylsilane columns to yield four glycosyl glycerides (Compounds 1-4), which were identified based on nuclear magnetic resonance, infrared, fast atom bombardment mass spectrometry, and gas chromatography/mass spectrometry data. Compounds 1-4 were evaluated for inhibition activity on NO production in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Results and conclusion: The glycosyl glycerides were identified to be (2S)-1-O-7(Z),10(Z),13(Z)-hexadecatrienoyl-3-O-${\beta}$-$\small{D}$-galactopyranosyl-sn-glycerol (1), (2S)-1-O-linolenoyl-3-O-${\beta}$-$\small{D}$-galactopyranosyl-sn-glycerol (2), (2S)-1-O-linolenoyl-2-O-linolenoyl-3-O-${\beta}$-$\small{D}$-galactopyranosyl-sn-glycerol (3), and 2(S)-1-O-linoleoyl-2-O-linoleoyl-3-O-${\beta}$-$\small{D}$-galactopyranosyl-sn-glycerol (4). Compounds 1 and 2 showed moderate inhibition activity on NO production in LPS-stimulated RAW264.7 cells [half maximal inhibitory concentration ($IC_{50}$): $63.8{\pm}6.4{\mu}M$ and $59.4{\pm}6.8{\mu}M$, respectively] without cytotoxicity at concentrations < $100{\mu}M$, whereas Compounds 3 and 4 showed good inhibition effect ($IC_{50}$: $7.7{\pm}0.6{\mu}M$ and $8.0{\pm}0.9{\mu}M$, respectively) without cytotoxicity at concentrations < $20{\mu}M$. All isolated compounds showed reduced messenger RNA (mRNA) expression of interleukin-$1{\beta}$ (IL-$1{\beta}$), IL-6, and tumor necrosis factor-${\alpha}$ in LPS-induced macrophage cells with strong inhibition of mRNA activity observed for Compounds 3 and 4.

Survey of mastitis management and incidence of mastitis in high somatic cell count of bulk milk at dairy farms in the Gyeongnam (경남지역의 체세포수 문제 목장에서의 젖소 유방염 관리실태 및 발생양상 조사)

  • Kim, Seong-Eun;Hah, Do-Yun;Jang, Eun-Hee;Kwon, Hee-Nyung;Jo, Seong-Suk;Kwon, Young-Taek;Park, Dong-Yeop;Lee, Kuk-Cheon;Kim, Jong-Shu
    • Korean Journal of Veterinary Service
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    • v.34 no.4
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    • pp.379-388
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    • 2011
  • Survey of mastitis management and incidence of mastitis in the Gyeongnam was started in May to September 2009 to solve mastitis problem statistically valid data for use in estimating mastitis management, isolation and antimicrobial drug susceptibility in 30 dairy farms having over 350,000/ml somatic cell count. In investigation on recognition of farmer about bovine mastitis, the ratio of understanding of differences between infectious and environmental origin, understanding of correlation between superbacteria and using indiscriminate, necessity of pathogen identification, and necessity of antimicrobial sensitivity tests were 80.0%, 73.3%, 33.3%, and 53.3%, respectively. In survey of mastitis management type, regular california mastitis test (CMT), conducting CMT test and empirical self-treatment, when detecting suspected cows, were 30.0%, 40.0%, and 46.7%, respectively. Checking and cleaning pulsators biweekly, cleaning vacuum system and replacing liners every 3~6 month, and getting milking system checked by engineers showed 80.0%, 76.7%, and 76.7% in the questionnaires, respectively. In recognition of farmer about milking hygiene for prevention of bovine mastitis, using individual towels, separated milking (milking order of cows), and teat-dipping disinfection after milking exhibited 13.3%, 86.7%, and 93.3%, respectively. In conclusion, through the questionnaires and laboratory test, we suggest that recognition of farmer about management and incidence of mastitis was very low, thus systemic educational program and public relations about mastitis management were need for dairy farmers.

Isolation, Identification, and Evaluation of Biocontrol Potentials of Rhizosphere Antagonists to Rhizoctonia solani (원예작물(園藝作物) 모잘록병(Rhizoctonia solani $K\"{u}hn$)의 발생(發生)에 관여하는 근권길항균(根圈拮抗菌)의 분리(分離), 동정(同定) 및 생물적(生物的) 방제(防除) 검토(檢討))

  • Kim, Hee-Kyu;Roh, Myung-Ju
    • Korean journal of applied entomology
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    • v.26 no.2 s.71
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    • pp.89-97
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    • 1987
  • Antagonistic microorganisms from rhizosphere soil were isolated, identified, and applied successfully as the biocontrol agents of damping-off caused by Rhizoctonia spp. Rhizosphere antagonists isolated from rhizosphere soil were identified as Trichoderma viride, T. harzianum, T. hamatum, T. polysporum, Gliocladium sp., Pseudomonas fluorescence, P. stutzeri, P. cepacia, Enterobacter sp., Serratia sp. and Erwinia herbicola. Of these, the most promising ones in vitro were T. virdie, T. harzianum, Gliocladium sp., Serratia sp., P. stutzeri, and P. cepacia. These above six antagonists were efficient in reducing disease incidence to $40{\sim}70%$ when the reselected rhizosphere antagonists preparations were applied to the soil at $10^6$ propagules per gram. Among six antagonists, T. viride was the most promising biocontrol agents against R. solani isolates in soil. The suppressive effect was more evident in steam-sterilized soil than in non-sterilized field soil.

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A STUDY OF NI-RESISTANT BACTERIA ISOLATED FROM GINGIVAL CREVICULAR FLUID ON THE PATIENTS WEARING NI-CR ALLOY PROSTHESIS (IN TERMS OF MOLECULAR BIOLOGICAL ASPECTS) (니켈-크롬 합금 보철물 주위 치은 열구내에서 발견된 니켈 내성 균주에 관한 분자생물학적 연구)

  • Chae Young-Ah;Woo Yi-Hyung;Kwon Kung-Rock
    • The Journal of Korean Academy of Prosthodontics
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    • v.41 no.2
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    • pp.207-222
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    • 2003
  • As a material of metal-ceramic prosthesis, nickel as a form of Ni-Cr alloy has been used for many dental prostheses in many cases. However, several problems in use of the alloy have been revealed (ex ; tissue stimulation, skin allergy, hypersensitivity cytotoxicity and carcinogenecity). Little is known about nickel with respect to the relationship between Ni-prosthesis and gaining of Niresistance in oral microorganisms. The present study was undertaken to check whether use of Ni-prosthesis leads to occurrence of Ni-resistant microorganisms. So this study may suggest the possible relationships between the oral microorganisms and nickel-resistance in oral environment. Bacteria were isolated from the gingival crevicular fluid on the patients wearing Ni-Cr prosthesis. The isolated bacteria were tested fir their Ni-resistance in nickel containing media at different concentration from 3mM to 110mM. E. coli HB101 was used as control. The Ni-resistant bacteria were isolated and biochemically identified. The Ni-resistant bacteria were tested several biochemical, molecular-biological tests. Performed tests were : measuring the growth curve, antibiotic test, growth ability test in liquid media, isolation of the chromosome and plasmid, digestion of DNA by restriction enzyme, electrophoresis of chromosome and plasmid DNA, identification of Ni-resistant genes by the DNA hybridization. The results were as follows 1) The bacteria isolated from gingival crevicular fluid on the patients wearing Ni-Cr alloy prosthesis showed nickel-resistance. 2) The isolated microorganisms grew at nickel containing media of high concentrations (60mM-110mM). 3) Based on the biochemical tests, the isolated microorganisms were identified as Enterococcus faecalis(13 cases), Klebsiella pneumoniae(1 case) and Enterobacter gergoviae(1 case). 4) Enterococcus faecalis expressed not only nickel resistance but also the multi-drug resistance to several antibiotics ; chloramphenicol, kanamicin, streptomycin, lincomycin, clindamycin, However, all strain showed the sensitivity against the tetracycline. 5) DNA hybridization result suggests that there is no homology between the previously known gene of nickel resistance in Klebsiella pneumoniae and chromosomal DNA of Enterococcus faecalis.

Characterization of Fusarium udum Causing Fusarium Wilt of Sunn Hemp in Korea (클로탈라리아 시들음병을 일으키는 Fusarium udum의 특성)

  • Choi, Hyo-Won;Hong, Sung Jun;Hong, Sung Kee;Lee, Young Kee;Kim, Jeomsoon
    • The Korean Journal of Mycology
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    • v.46 no.1
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    • pp.58-68
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    • 2018
  • Sunn hemp (Crotalaria juncea) is used as a nitrogen-fixing green manure in Korea to improve soil quality, reduce soil erosion, and suppress weeds and nematodes. In 2014, wilting sunn hemp plants were observed in green manure-cultivated fields in Wanju, Korea. Leaves of the infected plants began yellowing, starting with the lower leaves, eventually leading to their death. Moreover, a number of dark perithecia were observed on the wilting stems. Six isolates were obtained from these perithecia by single spore isolation. Based on their morphological characteristics, the isolates were identified as Fusarium udum (teleomorph: Gibberella indica). Macroconidia were slightly curved with almost hooked apical cell, and microconidia were formed on false heads by monophialides. Chlamydospores were produced abundantly in the hyphae, either singly or in clusters. To confirm the identification, multilocus sequence analysis was conducted using translation elongation factor 1 alpha (TEF), calmodulin (CAL), and histone 3 (HIS3). The sequences of TEF, CAL, and HIS3 showed 94.4~96.2%, 99.7%, and 99.6~99.8% similarity to the reference sequences of F. udum in NCBI GenBank, respectively. Pathogenicity was tested on sunn hemp and two soybean cultivars using the inoculation method of soil drenching with spore suspension. The wilting symptoms were observed only in sunn hemp and one cultivar of soybean (cv. Teagwang) after 14~21 days of inoculation. This is the first report of wilt disease in sunn hemp caused by Fusarium udum in Korea.

Isolation and Identification of Antioxidative Components from Bark of Rhus javanica Linne (붉나무 수피로부터 항산화 물질의 분리 및 동정)

  • Oh, Ji-Young;Choi, Ung;Kim, Yong-Suk;Shin, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.726-732
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    • 2003
  • The crude extracts from Rhus javanica Linne showed comparatively strong antioxidative activity in test oils. Antioxidative components were isolated and identified by column chromatography, thin layer chromatography, UV, and NMR. These antioxidative components were added to several oils to compare antioxidative activity with several commercial antioxidants, such as BHA, BHT, and tocopherol. After the sixth column chromatography, one fraction (R-18-9-3-2-4-2) was separated from chloroform layer of Rhus javanica Linne. The R-18-9-3-2-4-2 fraction was identified as methyl gallate by $^1H-NMR$ and $^{13}C-NMR$ and confirmed with methyl gallate standard as authentic. The R18-9-3-2-4-2 fraction from chloroform layer of Rhus javanica Linne showed stronger activity than that of the ${\alpha}-,\;{\delta}-tocopherol$, BHT, and BHA at the same concentration.

Isolation and Identification of Antioxidants from Makgeolli (막걸리로부터 항산화 화합물의 단리.정제 및 동정)

  • Wang, Seung-Jin;Lee, Hyoung-Jae;Cho, Jeong-Yong;Park, Keun-Hyung;Moon, Jae-Hak
    • Korean Journal of Food Science and Technology
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    • v.44 no.1
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    • pp.14-20
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    • 2012
  • The present study was carried out to isolate and identify of antioxidants from makgeolli. Makgeolli (3 L) was filtered and the residue was extracted by MeOH. The combined filtrates and MeOH extracts were successively solventfractionated by n-hexane, EtOAc, and BuOH. In the antioxidative activity against DPPH and $ABTS^+$ radicals of each fraction obtained after solvent-fractionation, EtOAc and BuOH layers showed higher activities than other fractions. Therefore, the two layers were respectively purified by column chromatography and HPLC. The isolated compounds were subjected to NMR and MS analyses and identified as 4-hydroxybenzaldehyde (1, 2.0 mg), 2-(4-hydroxyphenyl)ethanol (2, tyrosol, 15.3 mg), trans- and cis-ferulic acids (3 and 4, 1.2 mg), 1H-indole-3-ethanol (5, tryptophol, 3.4 mg), dimethyl succinate (6, 14.9 mg), succinic acid (7, 7.4 mg), and mono-methyl succinate (8, 7.8 mg). The presence of 1-5 in makgeolli have never before been reported.

Isolation and Identification of a Marine Bacterium, Pseudomonas sp. BK1 Producing Extracellular Enzymes Capable of Decomposing Multiple Complex Polysaccharides (복합 다당류 분해 효소들을 생산하는 해양미생물 Pseudomonas sp. BK1의 분리 및 특성)

  • Kim, Beom-Kyu;Jeon, Beong-Sam;Cha, Jae-Young;Park, Jeong-Won;Kim, Sam-Woong;Kim, Ji-Yoon;Park, Yong-Lark;Cho, Young-Su;Song, Jae-Young
    • Journal of Life Science
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    • v.13 no.6
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    • pp.871-878
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    • 2003
  • A marine bacterium (strain BKl) that produces extracellular enzymes capable of decomposing complex polysac-charides, such as agar, chitin, carboxymethylcellulose, xylan and mannan, was isolated from the marine red alga Porphyra dentata. Strain BKl was gram-negative, aerobic, catalase- and oxidase-positive, polarly flagellated bacilli that produce gelatinase and urease, but not decarboxylases. The G+C content of the DNA was 51.6 mol%. The major isoprenoid quinone component was identified as an ubiquinone-8, and the major cellular fatty acids were C16:0, C16:1 w6c and C18:1 w7c. Comparative 16S rRNA sequence analysis placed strain BK1 with members of the genus Pseudomonas. On the basis of phenotypic and genotypic data, the strain BK1 was shown to be a member of the subgroup of Pseudomonas, and named as Pseudomonas sp. BK1.

Isolation, Identification, and Characterization of Weissella Strains with High Ornithine Producing Capacity from Kimchi (김치로부터 오르니틴 생성능을 갖는 Weissella 속 균주의 분리, 동정 및 특성)

  • Yu, Jin-Ju;Park, Hyoung-Ju;Kim, Su-Gon;Oh, Suk-Heung
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.339-345
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    • 2009
  • Two lactic acid bacteria (LAB) with high ornithine-producing capacity were isolated from kimchi. Examination of the biochemical features using an API kit indicated that the strains belonged to the members of Weissella genus. They were gram positive, short rod-type bacteria, and able to grow anaerobically with $CO_2$ production. The isolates grew well on MRS broth at $25\sim37^{\circ}C$ and pH of 6.0~7.0. The optimum temperature and pH for growth are $30^{\circ}C$ and pH 6.5. The isolates fermented arabinose, ribose, xylose, glucose but not cellobiose, galactose, raffinose, or trehalsoe. The 16S rDNA sequences of isolates showed 99.6% and 99.7% homology with the Weissella koreensis S5623 16S rDNA (access no. AY035891). They were accordingly identified and named as Weissella koreensis OK1-4 and Weissella koreensis OK1-6, and could produce ornithine from MRS broth supplemented with 1% of arginine at a productivity of 27.01 and 31.41 mg/L/h, respectively. This is the first report on the production of ornithine by the genus Weissella isolated from kimchi.

Beneficial Effects of Kefir in Preventing and Treating Human Cancers (Kefir를 이용하여 다양한 Cancer 생성 예방 및 치료에 관한 연구)

  • Kim, Dong-Hyeon;Chon, Jung-Whan;Kim, Hyunsook;Lee, Soo-Kyung;Kim, Hong-Seok;Yim, Jin-Hyuk;Song, Kwang-Young;Kang, Il-Byung;Kim, Young-Ji;Jeong, Dana;Park, Jin-Hyeong;Jang, Ho-Seok;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.34 no.3
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    • pp.165-172
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    • 2016
  • Kefir, originating from Caucasus, is an acidic, alcoholic fermented milk product with little acidic taste and a creamy consistency. It is recognized in having beneficial effects infor the prevention and treatment of cancer. For example, Kefir has possesses a chemopreventative effect on carcinogenesis. There has recently been a strong focus on fermented milk foods containing a mixture of several functional organic substances and various probiotic microorganisms. Hence, the purpose of this review paper was to evaluate the scientific evidence for the effects of kefir on cancer prevention and treatment. Some of we analyzed and summarized data-relating to the effects of kefir on cancer. The cacers that kefir has an effect on are as follows: colon cancer, breast cancer, leukemia, sarcoma, skin cancer, gastric cancer. This review suggests that (1) kefir could be associated with cancer prevention, (2) kefir has beneficial effects in cancer treatment, and (3) kefir has various bioactive components including peptides, polysaccharides and sphingolipids, which contribute tofor itsthese anti-cancer properties. Furthermore, furthermore, studies were performed in order to obtain as to get the scientific evidence of kefir's anticancer activity: (1) improved protective effectiveness in vivo (human subjects or animal model), (2) isolation and identification of various bioactive components, and (3) mechanisms associated with beneficial effects.